Molecular Serotyping of<i>Klebsiella</i>Species Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

Brisse, Sylvain; Issenhuth-Jeanjean, Sylvie; Grimont, Patrick A. D.

doi:10.1128/jcm.42.8.3388-3398.2004

Cited by 56 publications

(78 citation statements)

References 39 publications

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“…1; see also File S3 in the supplemental material). This result is consistent with the similarity of the capsular polysaccharide (cps) clusters of these two serotypes (16). Serotype K4 is a hallmark of K. pneumoniae subsp.…”

Section: Resultssupporting

confidence: 79%

“…These isolates were selected to represent ST86 and ST380, which are associated with severe infections (1), and the nonvirulent ST14 clone (24). These K2 isolates also differed by their cps PCR-RFLP patterns, which were C patterns C2b to C2e (16). Nevertheless, the seven wzi sequences turned out to be totally identical (allele wzi-2).…”

Section: Resultsmentioning

confidence: 99%

“…However, K typing is technically difficult, and the production of the necessary reagents is complex. Molecular methods to deduce the K type from genomic sequences have been developed based on PCR restriction fragment length polymorphisms (PCR-RFLP) (16) or allele-specific PCR amplification (14,(17)(18)(19). Although the former method can be used to characterize most K. pneumoniae clinical isolates, it is technically complex.…”

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confidence: 99%

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wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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f Pathogens of the genus Klebsiella have been classified into distinct capsular (K) types for nearly a century. K typing of Klebsiella species still has important applications in epidemiology and clinical microbiology, but the serological method has strong practical limitations. Our objective was to evaluate the sequencing of wzi, a gene conserved in all capsular types of Klebsiella pneumoniae that codes for an outer membrane protein involved in capsule attachment to the cell surface, as a simple and rapid method for the prediction of K type. The sequencing of a 447-nucleotide region of wzi distinguished the K-type reference strains with only nine exceptions. A reference wzi sequence database was created by the inclusion of multiple strains representing K types associated with high virulence and multidrug resistance. A collection of 119 prospective clinical isolates of K. pneumoniae were then analyzed in parallel by wzi sequencing and classical K typing. Whereas K typing achieved typeability for 81% and discrimination for 94.4% of the isolates, these figures were 98.1% and 98.3%, respectively, for wzi sequencing. The prediction of K type once the wzi allele was known was 94%. wzi sequencing is a rapid and simple method for the determination of the K types of most K. pneumoniae clinical isolates.

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Section: Resultssupporting

confidence: 79%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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“…Therefore, PCR genotyping seems to be a more sensitive and specific way for detecting these serotypes (16). Furthermore, molecular serotyping is capable to determine a potential serotype of capsule-deficient isolates (7,18,19). We recommend that PCR analysis could be helpful in seroepidemiologic studies and early-tracing of contingent metastatic infections in patients with liver abscess (7,10,11,(15)(16)(17)(18)(19)(20).…”

Section: Discussionmentioning

confidence: 99%

Identification of Klebsiella pneumoniae K1 and K2 Capsular Types by PCR and Quellung Test

Feizabadi

Raji

Delfani

2013

Jundishapur J Microbiol

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Background: Klebseilla pneumoniae causes urinary tract infections, nosocomial pneumonia and intra-abdominal infections. Capsular antigens are considered to be the ultimate virulence determinants. Among 77 capsular serotypes of K. pneumoniae, serotypes K1 and K2 are the most virulent ones in humans. Objectives: We designed a PCR method for detection of capsular serotypes K1 and K2 of K. pneumoniae using genes cps cluster wzc and orf10 which are required for biosynthesis of capsular polysaccharides in K1 and K2 types, respectively. Materials and Methods: We collected 89 K. pneumoniae clinical isolates from the patients of Labafinejad Hospital located in Tehran. Clinical isolates were mostly chosen from urine. We used PCR technique to detect isolates possessing K1 and K2 capsular polysaccharides based on the wzc and orf10 genes which are required for biosynthesis of capsular polysaccharide in K1 and K2 types, respectively. The results of PCR were compared with those obtained by capsular serotyping using Quellung test. Results: Of 89 isolates of K. pneumoniae tested by both techniques, 10 (11.2%) belonged to K1 and 13 (14.6%) belonged to K2 serotypes, respectively. Conclusions: We found that these serotypes are probably important in clinical specimens. PCR was a simple and inexpensive tool for identification of K. pneumoniae clinical isolates of K1 and K2 geno-serotypes.

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“…Comparative studies were carried out to detect the correlation between different capsular serotype, mucoid or non-mucoid phenotype, and the presence of different genetic structures coding for virulence factors or harbored exclusively by virulent strains [10][11][12]. PCR-based subtractive hybridization technique was used to detect genetic sequences carried only by highly-virulent strains of K2 serotype of Klebsiella pneumoniae, and plasmid DNA analysis, amplified fragment length polymorphism (AFLP) pulsed-field gel electrophoresis (PFGE), restriction of the amplified capsular antigen gene cluster and multi locus sequence typing (http://www.mlst.net/) were used as molecular techniques to discriminate between strains [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Molecular Epidemiology and Virulence Characteristics of Klebsiella pneumoniae Strains Isolated from Hospital-Associated Infections

Damian¹,

Usein²,

Palade³

et al. 2009

TOEPIJ

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Abstract:Purpose: The present study aimed to confirm by classical and molecular laboratory methods hospitalassociated outbreaks due to virulent Klebsiella pneumoniae strains.Methods: Eighty three Klebsiella pneumoniae strains isolated from new-born patients, adults and hospital environment and devices in five hospital units, were analyzed for resistance to antibiotics, including last generation cephalosporins, sensitivity to bacteriophages and pulsed-field gel electrophoresis profiles in order to evaluate the epidemiological relatedness and their clonal spreading. Polymerase chain reaction targeting fur genes and several subtractive sequences (SL 002, SL 003, SL 019, SL 020, SL 021 and SL 025) was used for virulence assessment.Results: More than 50% of strains were resistant to third generation cephalosporins and among them 69% were extended spectrum beta-lactamase producers. Phage typing associated with pulse field gel electrophoresis documented the clonal dispersion of strains. Studying the distribution of virulence sequence, our results reveal that fur gene is present in all strains and among the subtractive sequences the most frequent is SL 020 followed by SL 019. None of the analyzed sequences are present in all clinical isolates and none of the bacterial strains carry all these sequences pointing out the heterogeneity of Klebsiella pneumoniae population. Conclusions:Phage typing method associated with pulse field gel electrophoresis typing and genetic profile for virulence indicated the occurrence of hospital associated-infections produced by Klebsiella pneumoniae strains. Moreover, the results reveal that virulence pattern could be used as a molecular marker in order to define strains which are involved in the process of the development of infectious diseases.

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Molecular Serotyping ofKlebsiellaSpecies Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

Cited by 56 publications

References 39 publications

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

Identification of Klebsiella pneumoniae K1 and K2 Capsular Types by PCR and Quellung Test

Molecular Epidemiology and Virulence Characteristics of Klebsiella pneumoniae Strains Isolated from Hospital-Associated Infections

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